Coding
Part:BBa_K4674001:Design
Designed by: Chun Ying Yu Group: iGEM23_CCU-Taiwan (2023-09-30)
mGL-4A-C7
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 16
Design Notes
The design of mGL-4A-C7
To efficiently label the CTCs captured by DNA tetrahedron, we first perform modeling and determine to link mGL protein and C7 dedocapeptide with 4 alaines (mGL-4A-C7). To expression the mGL-4A-C7 fusion protein, we design and cloned the composite par BBa_K4674010 into the pET15b. The fusion protein was induced by IPTG and purified by FPLC. Finally, to confirm whether the mGL-4A-C7 is functional, we perform the labeling experiments by SKOV3 cell line, which is the mimic of the captured CTC. (Due to the highly expression of FRα in SKOV3 cells)
Source
The mGL is derived from Aequorea victoria, while the C7 is a synthetic peptide.